Plate reader-based assays for measuring cell viability, neuroprotection and calcium in primary neuronal cultures Journal Article

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Authors: Burroughs, S. L.; Duncan, R. S.; Rayudu, P.; Kandula, P.; Payne, A. J.; Clark, J. L.; Koulen, P.; Kaja, S.
Article Title: Plate reader-based assays for measuring cell viability, neuroprotection and calcium in primary neuronal cultures
Abstract: Drug discovery and development efforts critically rely on cell-based assays for high-throughput screening. These assay systems mostly utilize immortalized cell lines, such as human embryonic kidney cells, and can provide information on cytotoxicity and cell viability, permeability and uptake of compounds as well as receptor pharmacology. While this approach has proven extremely useful for single-target pharmacology, there is an urgent need for neuropharmacological studies to screen novel drug candidates in a cellular environment resembles neurons in vivo more closely, in order to gain insight into the involvement of multiple signaling pathways. Primary cultured neuronal cells, such as cortical neurons, have long been used for basic research and low-throughput screening and assay development, and may thus be suitable candidates for the development of neuropharmacological high-throughput screening approaches. We here developed and optimized protocols for the use of primary cortical neuronal cells in high-throughput assays for neuropharmacology and neuroprotection, including calcium mobilization, cytotoxicity and viability as well as ion channel pharmacology. Our data show low inter-experimental variability and similar reproducibility as conventional cell line assays. We conclude that primary neuronal cultures provide a viable alternative to cell lines in high-throughput assay systems by providing a cellular environment more closely resembling physiological conditions in the central nervous system.
Journal Title: Journal of neuroscience methods
Volume: 203
Issue: 1
ISSN: 1872-678X; 0165-0270
Publisher: Elsevier Inc  
Journal Place: Netherlands
Date Published: 2012
Start Page: 141
End Page: 145
Language: ENG
DOI/URL:

10.1016/j.jneumeth.2011.09.007
Notes: LR: 20161019; CI: Copyright (c) 2011; GR: P01 AG022550/AG/NIA NIH HHS/United States; GR: S10 RR022570-01/RR/NCRR NIH HHS/United States; GR: P01 AG027956/AG/NIA NIH HHS/United States; GR: P01 AG010485-18/AG/NIA NIH HHS/United States; GR: AG010485/AG/NIA NIH HHS/United States; GR: R01 EY014227/EY/NEI NIH HHS/United States; GR: AG022550/AG/NIA NIH HHS/United States; GR: P01 AG010485/AG/NIA NIH HHS/United States; GR: P01 AG022550-09/AG/NIA NIH HHS/United States; GR: EY014227/EY/NEI NIH HHS/United States; GR: S10 RR027093-01/RR/NCRR NIH HHS/United States; GR: P01 AG027956-04/AG/NIA NIH HHS/United States; GR: RR022570/RR/NCRR NIH HHS/United States; GR: AG027956/AG/NIA NIH HHS/United States; GR: R01 EY014227-04/EY/NEI NIH HHS/United States; GR: S10 RR027093/RR/NCRR NIH HHS/United States; GR: S10 RR022570/RR/NCRR NIH HHS/United States; GR: RR027093/RR/NCRR NIH HHS/United States; JID: 7905558; 0 (Neuroprotective Agents); SY7Q814VUP (Calcium); NIHMS328819; OID: NLM: NIHMS328819; OID: NLM: PMC3221776; 2011/06/30 [received]; 2011/08/29 [revised]; 2011/09/07 [accepted]; ppublish